Egr2-CRE mice were stereotactically injected with AAVdj-DIO-eGFP (see Stereotactic surgery and virus injections ). Four weeks after the injection mice were deeply anesthetized with isoflurane, decapitated, and their brains were rapidly removed and briefly washed in PBS. Brains were placed in molds containing O.C.T embedding medium (Scigen Scientific Gardena, CA 90248 U.S.A) and snap-frozen on dry ice. Embedded brains were sectioned on a Leica CM1850 cryostat into 14µm sections, mounted onto SuperFrost Plus slides and kept at -80°C (see Histology ). Slides were processed according to ACD RNAscope fresh frozen tissue pretreatment and fluorescent multiplex assay manuals. All probes were purchased from Advanced Cell Diagnostics (RNAscope Probe – EGFP: Cat No. 400281, RNAscope Probe- Mm-Slc17a7-C2: Cat No. 416631-C2, RNAscope Probe- Mm-Gad2-C3: Cat No. 439371-C3). Slides were counterstained with DAPI for 30s, coverslipped with mounting medium (PermaFluor Aqueous Mounting Media, Thermo Scientific, Cat No. 94-TA-030-FM) and imaged using a high-speed fully-motorized multi-channel light microscope (Olympus IX-81) at 40x magnification. Data was analyzed using the CellProfiler v.2.1.1 (www.cellprofiler.org ) speckle counting pipeline with minor modifications [62 (link),63 (link)].
Permafluor aqueous mounting media
Manufactured by Thermo Fisher Scientific
Sourced in United States
PermaFluor Aqueous Mounting Media is a ready-to-use, water-based mounting medium designed for preparing permanent microscope slides. It is formulated to preserve fluorescent signals and provide long-term stability for fluorescently labeled specimens.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using permafluor aqueous mounting media
1
RNA Expression Mapping in Egr2-CRE Mice
2
Meridianin C-Induced Macropinocytosis Monitoring
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To monitor the functionality of meridianin C‐induced macropinocytosis (macropinosome formation/internalization), 0.25 × 105 YD‐10B cells/mL were seeded on coverslips and treated with meridianin C (1 μM) and/or FITC‐dextran (0.5 mg/mL) in the presence or absence of amiloride (4 mM) for 4 h. The cells were washed twice with PBS and mounted onto microscopic glass slides using Permafluor aqueous mounting media (Thermo Scientific, Waltham, MA, USA) media. Bright field and fluorescence were observed using a Zeiss AxioObserver.A1 inverted microscope (Carl Zeiss, Germany) and images acquired using Zen 2 software (Carl Zeiss). Fluorescent intensity was quantified using Image‐J software.
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